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Journal of virological methods1994; 46(2); 133-143; doi: 10.1016/0166-0934(94)90098-1

Detection of Borna disease virus RNA in naturally infected animals by a nested polymerase chain reaction.

Abstract: Borna disease virus in naturally infected horses, a donkey and sheep was detected for the first time by amplification of viral RNA using PCR. In contrast to a control group of healthy horses, brain tissue was positive by this assay in all animals with neurological symptoms. The use of a second round of PCR with nested primers following Southern hybridization confirmed the specificity and increased the sensitivity of the test. Comparison with conventional methods recommends this technique for monitoring of BDV infections at a molecular level.
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Publication Date: 1994-02-01 PubMed ID: 8188810DOI: 10.1016/0166-0934(94)90098-1Google Scholar: Lookup
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  • Comparative Study
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The study detected Borna disease virus in naturally infected animals using a polymerase chain reaction, demonstrating increased test specificity and sensitivity. This technique could be instrumental in monitoring BDV infections at the molecular level.

Introduction

  • The research focused on Borna disease virus (BDV), a pathogen that causes neurological symptoms in various animals, including horses, donkeys, and sheep.

    • Methodology

  • The researchers utilized a Polymerase Chain Reaction (PCR), a technique frequently used in molecular biology to make copies of a specific DNA segment, to detect BDV’s RNA.
  • A nested PCR was used, a modification of PCR specifically designed to improve the specificity of DNA amplification by reducing nonspecific amplification.

    • Results and Findings

  • For the first time, the researchers were able to detect BDV in naturally infected animals (horses, a donkey, and sheep) using this method. All the animals that tested positive for the virus through the PCR assay exhibited neurological symptoms. On the other hand, none of the brain tissues from the control group of healthy horses tested positive.
  • The researchers employed a second round of PCR to confirm the initial results. This was done with nested primers that target a smaller internal region of the DNA or RNA to be amplified.
  • The researchers substantiated the PCR results via Southern hybridization, a method used to check for the existence of a particular DNA sequence within a DNA sample.

    • Conclusion

  • The combination of the two PCR rounds and Southern hybridization not only confirmed the specificity of the test but also increased its sensitivity, making it a reliable tool for BDV detection.
  • The study suggests that this method is more effective compared to conventional techniques, recommending it for future BDV infection monitoring at the molecular level.

    • Cite This Article

    APA
    Zimmermann W, Dürrwald R, Ludwig H. (1994). Detection of Borna disease virus RNA in naturally infected animals by a nested polymerase chain reaction. J Virol Methods, 46(2), 133-143. https://doi.org/10.1016/0166-0934(94)90098-1

    Publication

    Journal of virological methods
    ISSN: 0166-0934
    NlmUniqueID: 8005839
    Country: Netherlands
    Language: English
    Volume: 46
    Issue: 2
    Pages: 133-143

    Researcher Affiliations

    Zimmermann, W
    • Institut für Virologie, Freie Universität Berlin, Germany.
    Dürrwald, R
      Ludwig, H

        MeSH Terms

        • Animals
        • Base Sequence
        • Borna Disease / microbiology
        • Borna disease virus / isolation & purification
        • Brain / microbiology
        • Horse Diseases / microbiology
        • Horses
        • Molecular Sequence Data
        • Polymerase Chain Reaction / methods
        • Polymerase Chain Reaction / veterinary
        • RNA, Viral / genetics
        • RNA, Viral / isolation & purification
        • Sensitivity and Specificity
        • Sheep
        • Sheep Diseases / microbiology
        • Virology / methods

        Citations

        This article has been cited 7 times.
        1. Schindler AR, Vögtlin A, Hilbe M, Puorger M, Zlinszky K, Ackermann M, Ehrensperger F. Reverse transcription real-time PCR assays for detection and quantification of Borna disease virus in diseased hosts. Mol Cell Probes 2007 Feb;21(1):47-55.
          doi: 10.1016/j.mcp.2006.08.001pubmed: 17014984google scholar: lookup
        2. Hornig M, Briese T, Lipkin WI. Borna disease virus. J Neurovirol 2003 Apr;9(2):259-73.
          doi: 10.1080/13550280390194064pubmed: 12707857google scholar: lookup
        3. Gonzalez-Dunia D, Sauder C, de la Torre JC. Borna disease virus and the brain. Brain Res Bull 1997;44(6):647-64.
          doi: 10.1016/s0361-9230(97)00276-1pubmed: 9421127google scholar: lookup
        4. Kishi M, Nakaya T, Nakamura Y, Kakinuma M, Takahashi TA, Sekiguchi S, Uchikawa M, Tadokoro K, Ikeda K, Ikuta K. Prevalence of Borna disease virus RNA in peripheral blood mononuclear cells from blood donors. Med Microbiol Immunol 1995 Oct;184(3):135-8.
          doi: 10.1007/BF00224350pubmed: 8577314google scholar: lookup
        5. Pfeffer M, Wiedmann M, Batt CA. Applications of DNA amplification techniques in veterinary diagnostics. Vet Res Commun 1995;19(5):375-407.
          doi: 10.1007/BF01839319pubmed: 8560754google scholar: lookup
        6. Kishi M, Arimura Y, Ikuta K, Shoya Y, Lai PK, Kakinuma M. Sequence variability of Borna disease virus open reading frame II found in human peripheral blood mononuclear cells. J Virol 1996 Jan;70(1):635-40.
          doi: 10.1128/JVI.70.1.635-640.1996pubmed: 8523585google scholar: lookup
        7. Zimmermann W, Breter H, Rudolph M, Ludwig H. Borna disease virus: immunoelectron microscopic characterization of cell-free virus and further information about the genome. J Virol 1994 Oct;68(10):6755-8.
          doi: 10.1128/JVI.68.10.6755-6758.1994pubmed: 8084008google scholar: lookup
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